Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

( Xing Jia Shen ),( Yong Zhu Yi ),( Shun Ming Tang ),( Zhi Fang Zhang ),( Yi Ren Li ),( Jia Lu He ),( Xiang Fu Wu ) 한국잠사학회 2007 International Journal of Industrial Entomology Vol.8 No.2

Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (luc) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0∼2.0㎍/㎖ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5∼2.0㎍/㎖ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

A Hierarchical Genetic Algorithm based RBF Neural Network Approach for Modelling of Electrohydraulic System

Xing Zong-yi,Zhang Yuan,Qin Yong,Jia Li-min,Wu Ying-ying 제어로봇시스템학회 2009 제어로봇시스템학회 국제학술대회 논문집 Vol.2009 No.8

The paper presents an approach to model the electrohydraulic system of a certain mine-sweeping weapon using the Radial Basis Function (RBF) neural networks. In order to obtain accurate and simple RBF neural networks efficiently, a hierarchical genetic algorithm (HGA) is used to train the neural networks, in which the number of hidden units and the parameters of centers are optimized by the HGA simultaneously. The spread factors and the weights of the neural networks are calculated by the linear algebra methods for relieving computational burden. The proposed algorithm is applied to the modelling of the electrohydraulic system, and the results clearly indicate that the obtained RBF neural network can model the hydraulic system satisfactorily. The comparison results also show that the proposed algorithm performs better than the traditional methods.

Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

Wu, Xiang-Mei,Liu, Xing,Jiao, Qing-Fang,Fu, Shao-Yue,Bu, You-Quan,Song, Fang-Zhou,Yi, Fa-Ping Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

Diversity and Active Mechanism of Fengycin-Type Cyclopeptides from Bacillus subtilis XF-1 Against Plasmodiophora brassicae

( Xing Yu Li ),( Zi Chao Mao ),( Yue Hu Wang ),( Yi Xing Wu ),( Yue Qiu He ),( Chun Lin Long ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.3

Bacillus subtilis XF-1, a strain with demonstrated ability to control clubroot disease caused by Plasmodiophora brassicae, was studied to elucidate its mechanism of antifungal activity against P. brassicae. Fengycin-type cyclopeptides (FTCPs), a well-known class of compounds with strong fungitoxic activity, were purified by acid precipitation, methanol extraction, and chromatographic separation. Eight homologs of fengycin, seven homologs of dehydroxyfengycin, and six unknown FTCPs were characterized with LC/ESI-MS, LC/ESI-MS/MS, and NMR. FTCPs (250μg/ml) were used to treat the resting spores of P. brassicae (107/ml) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be cleaved by the FTCPs of B. subtilis XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol.

Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

Xing-Jia Shen,Yong-Zhu Yi,Shun-Ming Tang,Zhi-Fang Zhang,Yi-Ren Li,Jia-Lu He,Xiang-Fu Wu 한국잠사학회 2004 International Journal of Industrial Entomology Vol.8 No.2

Bombyx mori nucleopolyhedrovirus (BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegt159- luc with luciferase (luc) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 ~ 2.0 mg/ml can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5 ~ 2.0 mg/ml increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Betulin Targets Lipin1/2-Meidated P2X7 Receptor as a Therapeutic Approach to Attenuate Lipid Accumulation and Metaflammation

( Jia-yi Dou ),( Yu-chen Jiang ),( Zhong-he Hu ),( Kun-chen Yao ),( Ming-hui Yuan ),( Xiao-xue Bao ),( Mei-jie Zhou ),( Yue Liu ),( Zhao-xu Li ),( Li-hua Lian ),( Ji-xing Nan ),( Yan-ling Wu ) 한국응용약물학회 2022 Biomolecules & Therapeutics(구 응용약물학회지) Vol.30 No.3

The present study focused on the potential mechanism of betulin (BT), a pentacyclic triterpenoid isolated from the bark of white birch (Betula pubescens), against chronic alcohol-induced lipid accumulation and metaflammation. AML-12 and RAW 264.7 cells were administered ethanol (EtOH), lipopolysaccharide (LPS) or BT. Male C57BL/6 mice were fed Lieber-DeCarli liquid diets containing 5% EtOH for 4 weeks, followed by single EtOH gavage on the last day and simultaneous treatment with BT (20 or 50 mg/kg) by oral gavage once per day. In vitro, MTT showed that 0-25 mM EtOH and 0-25 μM BT had no toxic effect on AML-12 cells. BT could regulate sterolregulatory-element-binding protein 1 (SREBP1), lipin1/2, P2X7 receptor (P2X7r) and NOD-like receptor family, pyrin domains-containing protein 3 (NLRP3) expressions again EtOH-stimulation. Oil Red O staining also indicated that BT significantly reduced lipid accumulation in EtOH-stimulated AML-12 cells. Lipin1/2 deficiency indicated that BT might mediate lipin1/2 to regulate SREBP1 and P2X7r expression and further alleviate lipid accumulation and inflammation. In vivo, BT significantly alleviated histopathological changes, reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and triglyceride (TG) levels, and regulated lipin1/2, SREBP1, peroxisome proliferator activated receptor α/γ (PPARα/γ) and PGC-1α expression compared with the EtOH group. BT reduced the secretion of inflammatory factors and blocked the P2X7r-NLRP3 signaling pathway. Collectively, BT attenuated lipid accumulation and metaflammation by regulating the lipin1/2-mediated P2X7r signaling pathway.

Confinement coefficient of concrete-filled square stainless steel tubular stub columns

Fa-Xing Ding,Yi-xiang Yin,Liping Wang,Yujie Yu,Liang Luo,Zhi-wu Yu 국제구조공학회 2019 Steel and Composite Structures, An International J Vol.30 No.4

The objective of this paper is to investigate the confinement coefficient of concrete-filled square stainless steel tubular (CFSSST) stub columns under axial loading. A fine finite 3D solid element model was established, which utilized a constitutive model of stainless steel considering the strain-hardening characteristics and a triaxial plastic-damage constitutive model of concrete with features of the parameter certainty under axial compression. The finite element analysis results revealed that the increased ultimate bearing capacity of CFSSST stub columns compared with their carbon steel counterparts was mainly due to that the composite action of CFSSST stub columns is stronger than that of carbon steel counterparts. A further parametric study was carried out based on the verified model, and it was found that the stress contribution of the stainless steel tube is higher than the carbon steel tube. The stress nephogram was simplified reasonably in accordance with the limit state of core concrete and a theoretical formula was proposed to estimate the ultimate bearing capacity of square CFSSST stub columns using superposition method. The predicted results showed satisfactory agreement with both the experimental and FE results. Finally, the comparisons of the experimental and predicted results using the proposed formula and the existing codes were illustrated.

Reference gene selection for transcriptional profiling by RT-qPCR in the 28-spotted larger potato ladybird

Zhang Yu-Xing,Tan Qiao,Shen Chen-Hui,Wu Jian-Jian,Wu Yi-Kuan,Li Wen-Ze,Jin Lin,Li Guo-Qing 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.2

Henosepilachna vigintioctomaculata is one of the most serious defoliates attacking potatoes. However, studies on functional genes have greatly been limited due to the insufficiency of effective and stable endogenous references to normalize RT-qPCR data. In this report, nine housekeeping genes (RPL4, RPL6, RPL13, RPL32, RPS18, ACT, EF1α, GAPDH and α-TUB) involved in different biological processes were selected. Their expression levels under diverse experimental conditions including developmental stages, tissues, temperatures and host plants were determined using RT-qPCR technology. The tested candidate genes were comprehensively ranked based on five alternative stability analysis methods (Ct value, geNorm, NormFinder, BestKeeper and ReFinder). The results revealed that the optimal internal reference genes varied under different experimental conditions. Any gene pair among the five candidates (RPL4, RPL13, RPL32, RPS18 and EF1α) was a suitable reference gene set under different temperatures and on different host plants. A combination of RPL6 and RPL13 was recommended as the best reference gene set across different developmental stages. A pair of RPS18 and EF1α was ranked as the optimal reference gene combination within different tissues. The most suitable reference genes were RPS18 and RPL13 under four different experimental conditions. Our findings not only establish an accurate and reliable normalization of RT-qPCR data, but also lay a solid foundation for further functional gene researches in H. vigintioctomaculata

Study on Interpretable Fuzzy Classification System Based on Neural Networks

Qin Yong,Xing Zong-yi,Jia Li-min,Wu Ying-ying 제어로봇시스템학회 2009 제어로봇시스템학회 국제학술대회 논문집 Vol.2009 No.8

This paper describes a comprehensive method to construct fuzzy classification system considering bothprecision and interpretability. Fuzzy classification system, initialized by modified Gath-Geva fuzzy clustering algorithm, is transformed into neural network. After training the neural network, fuzzy sets similarity measure is adopt to mergeredundant fuzzy sets to improve interpretability, and a constraint genetic algorithm is applied to improve precision. The simulation result on Iris data problem demonstrates the effectiveness of the proposed method

Prevalence of Spina Bifida Occulta and Its Relationship With Overactive Bladder in Middle-Aged and Elderly Chinese People

Jun Wei Wu,Yu Rong Xing,Yi Bo Wen,Tian Fang Li,Quan De Feng,Xiao Ping Shang,Yun Long Li,Jin Jin Feng,Xin Xin Wang,Rong Qun Zhai,Xiang Fei He,Tao Chen,Xin Jian Liu,Jian Guo Wen 대한배뇨장애요실금학회 2016 International Neurourology Journal Vol.20 No.2

Purpose: To investigate the prevalence of spina bifida occulta (SBO) and its relationship with the presence of overactive bladder (OAB) in middle-aged and elderly people in China. Methods: A cross-sectional community-based survey was carried out at 7 communities in Zhengzhou City, China from December 15, 2013 to June 10, 2014, where residents aged over 40 years were randomly selected to participate. All of the participants underwent lumbosacral radiographic analysis and relevant laboratory tests. A questionnaire including basic information, past medical history and present illness, and the OAB symptom score was filled out by all participants. Chi-square tests and logistic regression were used for data analysis with a P-value of <0.05 denoting statistical significance. Results: A total of 1,061 subjects were qualified for the final statistical analysis (58.8±11.7 years; male, 471 [44.4%]; female, 590 [55.6%]). The overall prevalence of SBO was 15.1% (160 of 1,061): 18.3% (86 of 471) in men and 12.5% (74 of 590) in women. Among these subjects, 13.7% (145 of 1,061) had OAB: 13.2% (62 of 471) in men and 14.1% (83 of 590) in women. The results of logistic regression showed that age, SBO, history of cerebral infarction (HCI), and constipation were risk factors for OAB (P<0.05), while sex, history of childhood enuresis (HCE), body mass index (BMI), and diabetes mellitus (DM) were not (P>0.05). In men, age, SBO, and constipation were risk factors for OAB (P<0.05), while HCE, BMI, DM, HCI, and benign prostate hyperplasia were not (P>0.05). In women, age, SBO, and HCI were risk factors for OAB (P<0.05), while HCE, BMI, DM, vaginal delivery, and constipation were not (P>0.05). Conclusions: The prevalence of SBO is high and it is related to OAB in middle-aged and elderly people in China.